Development of a human whole blood assay for prediction of cytokine release similar to anti-CD28 superagonists using multiplex cytokine and hierarchical cluster analysis

► An assay for detection of anti-CD28 mediated cytokine release is described. ► We tested 8 marketed mAbs for cytokine production similar to anti-CD28 superagonists. ► 9 cytokines analyzed in addition to the previously published IL-6, INF-γ, and TNF-α. ► Multivariate hierarchical cluster analysis was used to analyze the data set. ► Results show that the test mAbs are distinct form anti-CD28 superagonists.